The use of hematoporphyrin and its acetylated derivative mixture hematoporphyrin derivative (HPD) systemically, combined with irradiation, for the detection and treatment of malignant cells has, by this time, some considerable history. HPD is a mixture of porphyrins including hematoporphyrin itself, hydroxyethyl vinyl deuteroporphyrin, protoporphyrin, and dihematoporphyrin ethers. (See, e.g., "Porphyrin Photosensitization," Kessel, D., et al., eds. (1983) Plenum Press.)
HPD seems "naturally" capable of localizing in malignant cells. When irradiated, it has two properties which make it useful. First, when irradiated with ultraviolet or visible light, it is capable of fluorescence, and thus is useful in diagnostic methods related to detection of malignancy (see, for example, Kessel, D., et al. (supra); Gregory, H. B., Jr., et al., Ann Surg (1968) 167:827-829). Also important is the capacity of HPD, when irradiated with visible light, to exhibit a cytotoxic effect on the cells or other tissue in which it is localized (see, for example, Diamond, I., et al., Lancet (1972) 2:1175-1177; Dougherty, T. J., et al., Cancer Research (1978) 38:2628-2635; Dougherty, T. J., et al., "The Science of Photo Medicine" (1982) J. D. Regan & J. A. Parrish, eds., pp. 625-638; Dougherty, T. J., et al., "Cancer: Principles and Practice of Oncology" (1982) V. T. DeVita Jr., et al., eds., pp. 1836-1844). Although it has not been definitively established, the effect of HPD in killing cells seems to be due to the formation of singlet oxygen upon irradiation (Weishaupt, K. R., et al., Cancer Research (1976) 36:2326-2329). Several mechanisms for this effect have been proposed, and it has been shown that the active ingredient(s) in HPD which mediates the cytotoxic effect of visible light irradiation is the mixture of dihematoporphyrin ethers (DHE) (Dougherty, T. J., et al., "Porphyrin Localization and Treatment of Tumors" (1984) pp. 301-314; Dougherty, T. J., CRC Critical Reviews in Oncology/Hematology (1984) 2:83-116).
A purified form of the active component(s) of HPD is obtained by adjustment of pH to cause aggregation and recovery of the aggregate, as disclosed in U.S. Pat. Nos. 4,649,151, 4,866,168, 4,889,129 and 4,932,934. The purified form called DHE in the patent, is marketed under the trademark Photofrin.RTM. II and has been used in a manner completely analogous to HPD.
In addition to in vivo therapeutic and diagnostic protocols for tumors as described in the above-cited patent, the porphyrins, including HPD and its more purified derivatives, can be used in other in vivo and in vitro applications. For example, photosensitizers are useful in the detection and treatment of atherosclerotic plaques as described in U.S. Pat. Nos. 4,512,762 and 4,577,636. U.S. Pat. Nos. 4,500,507 and 4,485,806 describe the use of radiolabeled porphyrin compounds, including HPD, for tumor imaging. U.S. Pat. No. 4,753,958 to the University of California describes the use of topical application of porphyrin sensitizers for diagnosis and treatment of skin diseases. U.S. Pat. Nos. 4,748,120 and 4,878,891 describe the use of photosensitizers in the treatment of whole blood or blood components. Photochemical decontamination treatment of blood and components is also described in U.S. Pat. No. 4,727,027 where the photosensitizer is furocoumarin and its derivatives. In addition, viruses are inactivated in therapeutic protein compositions in vitro as disclosed in U.S. Pat. No. 4,268,947.
The successful treatment of AIDS-related oral Kaposi's Sarcoma with the related photosensitizer Photofrin.RTM. II porfimer sodium was described by Schweitzer, V. G., at al., Otolaryngology--Head and Neck Surgery (1990) 102:639-649. It is expected that the modified porphyrins of the present invention will also be effective in regard to this indication and that the sole related side effect--hypersensitivity to sunlight--will be avoided due to the lower dose levels required for the invention compounds.
While the treatment of tumors and other undesirable targets with HPD relies on the intrinsic ability of HPD to localize in malignant or other relevant cells or targets, a considerable improvement and refinement in specificity has been achieved by conjugating the hematoporphyrin to target-specific antibodies. For example, when hematoporphyrin was coupled to monoclonal antibodies directed to a murine myosarcoma cell line M1, administration of anti-M1 hematoporphyrin-conjugates to tumor-bearing animals followed by exposure to incandescent light resulted in the suppression of M1 growth (Mew, D., et al., J Immunol (1983) 130:1473-1477). In additional work, hematoporphyrin was conjugated to a monoclonal antibody specific to an antigen associated with a human leukemia (CAMAL) and the conjugates were shown to mediate the irradiation-induced killing of leukemic cells specifically, in vitro (Mew, D., et al., Cancer Research (1985) 45:4380-4386). Conjugation of the related compound chlorin e.sub.6 to anti-T cell Mab has also been reported (Oseroff, A. R., et al., Proc Natl Acad Sci USA (1986) 83:8744-8748).
While the conjugation of hematoporphyrin to immunoglobulins specific for targeted cells refines the ability of the hematoporphyrin to home to the desired cells or tissue, this still does not solve another problem ancillary to this general therapeutic approach, namely that the wavelength for irradiation required to activate the hematoporphyrin or HPD, which is in the range of 630 nanometers, is also an energy which is readily absorbed by the endogenous porphyrins and other natural chromophores in the blood and other tissues. Therefore, relatively large amounts of the hematoporphyrin or HPD must be administered, often resulting in oversensitization of the patient to light in general. It would be desirable to administer compounds to mediate the effects of irradiation in a lower amount and with higher clearance rates, thus avoiding the problems of hypersensitivity exhibited nonspecifically throughout the subject organism. The activity of certain of these compounds was described in a paper by Richter, A. M., et al., in J Natl Cancer Inst (1987) 79:1327-1332, mailed to subscribers on Jan. 19, 1988. The invention is directed to the use of such compounds.